Quality Assessment of Sitopaladi Churna Using High-Performance Liquid Chromatography Coupled with Multivariate Analysis.

"Sitopaladi churna," a well-known formulation of Ayurveda, is prescribed to treat the disease like Bronchitis, Pneumonia, allergic conditions, viral infection of the respiratory tract and as a natural antioxidant. A novel method based on reversed-phase high-performance liquid chromatography (RP-HPLC) coupled to photodiode array detector was established and validated for sitopaladi churna. Here, HPLC fingerprints data obtained for 28 samples including standard sample were then treated to chemometric analysis like principal component analysis and hierarchical clustering analysis for further analysis to evaluate the differences in market samples of sitopaladi churna. Additionally, one major marker compound, piperine was quantified and it also facilitated for relative retention time. The simulative mean spectrum was also generated. The validation results showed that the developed method was simple, precise and stable. Thus, the developed chromatographic method adjoined with multivariate analysis can be used as an efficient and practical approach for quality assessment of sitopaladi churna.

Development and Validation of a High-Performance Thin-Layer Chromatographic (HPTLC) Method for Simultaneous Quantification of Reserpine, Atropine, and Piperine in Sarpagandha Ghanvati, a Classical Ayurvedic Preparation.

Background: Anxiety disorders are the most common of emotional disorders, affecting more than 20 million people annually. Sarpagandha Ghanvati is a classical Ayurvedic polyherbal formulation prescribed in conditions of insomnia, hysteria, and is used as an anxiolytic agent. Standardization and quality control are the two major issues that need to be addressed for herbal formulations, especially those containing multiple herbal ingredients. Objective: An HPTLC method was developed for the simultaneous quantification of reserpine, atropine, and piperine from Sarpagandha Ghanvati containing Rauwolfia serpentine (root), Hyoscyamus niger (seed), and Piper longum (root and stem). Methods: The marker compounds were effectively resolved on a silica gel G TLC plate using toluene-ethyl acetate-diethyl amine (7+2+1, v/v) as the mobile phase. The detected wavelengths for reserpine, atropine, and piperine were 269, 220, and 254 nm, respectively. The method was validated as per the International Conference on Harmonization guidelines. Results: R. serpentine roots contained 0.82% w/w of reserpine. Atropine content in the seeds of H. niger was found to be 0.004% w/w, whereas P. longum roots were found to contain 0.508% of piperine. The method was found to be accurate, which was evident from 98.93, 99.46, and 99.10% recovery of reserpine, atropine, and piperine, respectively, when the respective herbs were spiked with them. By the developed HPTLC method, 1.0 g of Sarpagandha Ghanvati was found to contain 4.94, 0.049, and 0.318 mg of reserpine, atropine, and piperine, respectively. The recoveries of these three markers from the formulation were found to be 90.32, 92.45, and 89.97%, respectively. Conclusions: The developed method can be successfully used for simultaneous estimation of these marker compounds and for the quality control of the classical Ayurvedic formulation Sarpagandha Ghanvati. Highlights: This works describes effects of extraction solvents on the quantities of marker compounds in the formulations. It also suggests a simple and reliable HPTLC method for simultaneous quantification of three different marker compounds from a poly-herbal formulation.

Evaluation of free radical scavenging properties of two classical polyherbal formulations.

Two polyherbal formulations of Ayurveda viz., Chandraprabha Vati and Maha yogaraja Guggulu were evaluated for their free radical scavenging properties. Methanolic extracts of the formulations were studied in four different in vitro and ex vivo models. Total phenolic content of Chandraprabha Vati and Maha yogaraja Guggulu was found to be 5.24% and 10.74% respectively. Methanolic extracts of the formulations were good scavengers of all the radicals but there was a difference in the activity of the two formulations in different models. Chandraprabha Vati was a good scavenger of superoxide radical and Maha yogaraja Guggulu was efficient in scavenging nitric oxide (NO), while both inhibited lipid peroxidation efficiently. Free radical scavenging activity of the different extracts can be attributed to the presence of various chemical components including phenolics.

Development and validation of a thin-layer chromatography-densitometric method for the quantitation of alliin from garlic (Allium sativum) and its formulations.

A selective, precise, and accurate high-performance thin-layer chromatographic (HPTLC) method has been proposed for the analysis of garlic and its formulations for their alliin content. The method involves densitometric evaluation of alliin after resolving it by HPTLC on silica gel plates with n-butanol-acetic acid-water (6 + 2 + 2, v/v) as the mobile phase. For densitometric evaluation, peak areas were recorded at 540 nm after derivatizing the resolved bands with ninhydrin reagent. The relation between the concentration of alliin and corresponding peak areas was found to be linear within the range of 250 to 1500 ng/spot. The method was validated for precision (interday and intraday), repeatability, and accuracy. Mean recovery was 98.36%. The method was applied for the quantitation of alliin in bulbs of Allium sativum Linn. (garlic) and its formulations. The proposed TLC method was found to be precise, specific, sensitive, and accurate and can be used for routine quality control of garlic and its formulations.